Ramírez-Hernández C, Hernández-Vidal G, Wong-González A, Gutiérrez-Ornelas E, Ackermann M R, Ramírez-Romero R
Departamento de Patología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma de Nuevo León, Av. Lázaro Cárdenas 4600, Unidad Mederos, Monterrey, N.L. 64930, México.
Inflamm Res. 2004 Mar;53(3):107-10. doi: 10.1007/s00011-003-1229-x. Epub 2004 Feb 16.
To quantify the number of mast cells in the skin of rabbits during initiation and progression of the local Shwartzman reaction.
Thirty New Zealand rabbits were divided in two groups (n = 15/group). One group was subjected to the Shwartzman reaction and the other group served as control. Subsequently, animals were further subdivided in six groups of five animals each according to time of euthanasia.
The local Shwartzman reaction was induced by two inoculations of Salmonella typhimurium lipopolysaccharide. Preparatory inoculation was given intradermally and, 24 h later, the provocative injection was administered intravenously. Controls were subjected to the same procedure but received saline. After provocative injection animals were killed at 1, 8, and 15 days.
Skin samples were fixed in Carnoy's solution and mast cells were identified employing a low pH toluidine blue stain. Numbers of mast cells were determined per square millimetre and, subsequently, those cells degranulated were identified and quantified to obtain absolute values. A Student's t test was initially used to compare Shwartzman versus controls at each time point. Subsequently, an ANOVA test employing a factorial experiment was used to assess a possible interaction between time of euthanasia and treatments.
The values were transformed (natural logarithms) for appropriate statistical comparisons. Independent comparisons at each time point showed that Shwartzman groups had higher numbers of mast cells than controls at 1 and 8 days, but not at 15 days (5.71 +/- 1.00 Vs. 2.40 +/- 0.58, P < 0.005; 3.77 +/- 0.90 Vs. 2.33 +/- 0.56, P < 0. 025, and 2.61 +/- 0.25 Vs. 2.39 +/- 0.39, P > 0.05, respectively). Degranulated cells were numerous in Shwartzman groups, particularly at day 1 (3.48 +/- 0.78) and less obvious at day 8 (0.72 +/- 0.50), but were scarce by day 15 (-0.67 +/- 0.99) as well as controls (-0.68 +/- 0.91). The factorial experiment demonstrated that the Shwartzman reaction and time of euthanasia were independently significant (P < 0.005) but their interaction at day 1 was the major contributor (P < 0.005). Tukey's w pairwise comparisons of means confirmed that the Shwartzman group killed at day 1 was significantly different from the others (P < 0.05).
Mast cells significantly increase in the early stages of the local Shwartzman reaction. Thus, mast cells are a highly dynamic cell population, which have a prominent role during the acute phase of this lipopolysaccharide-induced inflammatory reaction but not during healing.
量化局部施瓦茨曼反应起始和进展过程中兔皮肤内肥大细胞的数量。
30只新西兰兔分为两组（每组n = 15只）。一组进行施瓦茨曼反应，另一组作为对照。随后，根据安乐死时间将动物进一步分为六组，每组五只。
通过两次接种鼠伤寒沙门氏菌脂多糖诱导局部施瓦茨曼反应。预先接种采用皮内注射，24小时后静脉注射激发剂。对照组接受相同操作，但注射生理盐水。激发注射后，在第1、8和15天处死动物。
皮肤样本用卡诺氏固定液固定，采用低pH甲苯胺蓝染色鉴定肥大细胞。每平方毫米测定肥大细胞数量，随后鉴定并量化脱颗粒细胞以获得绝对值。最初使用学生t检验在每个时间点比较施瓦茨曼组与对照组。随后，采用析因实验的方差分析来评估安乐死时间与处理之间可能的相互作用。
为进行适当的统计比较，对数值进行了转换（自然对数）。每个时间点的独立比较显示，施瓦茨曼组在第1天和第8天的肥大细胞数量高于对照组，但在第15天并非如此（分别为5.71±1.00对2.40±0.58，P < 0.005；3.77±0.90对2.33±0.56，P < 0.025，以及2.61±0.25对2.39±0.39，P > 0.05）。施瓦茨曼组中脱颗粒细胞数量众多，尤其是在第1天（3.48±0.78），在第8天不太明显（0.72±0.50），但到第15天（-0.67±0.99）以及对照组（-0.68±0.91）时则很少。析因实验表明，施瓦茨曼反应和安乐死时间各自具有显著意义（P < 0.005），但它们在第1天的相互作用是主要因素（P < 0.005）。Tukey's w均值两两比较证实，在第1天处死的施瓦茨曼组与其他组有显著差异（P < 0.05）。
在局部施瓦茨曼反应的早期阶段，肥大细胞显著增加。因此，肥大细胞是一个高度动态的细胞群体，在这种脂多糖诱导的炎症反应急性期起重要作用，但在愈合过程中并非如此。
